1,2-methylene-b-norsteroids



United States Patent 3,449,434 1,Z-METHYLENE-B-N()RSTEROIDS Kenneth G.Holden, Stratford, N.J., assignor to Smith Kline & French Laboratories,Philadelphia, Pa., a corporation of Pennsylvania N0 Drawing. Filed Sept.21, 1966, Ser. No. 580,901 Int. Cl. 'C07c 49/44; C12b 1/00 US. Cl.260-586 4 Claims ABSTRACT OF THE DISCLOSURE1a,Za-methyIene-B-norandrostenes, unsubstituted at the 6-position, orsubstituted with a 6,6-ethylene or 6-methy1 group, are prepared byreaction of the corresponding 1,4- androstadienes with dimethylsulfoxonium methylide. The products possess antiandrogenic activity.

This invention relates to steroid compounds having antiandrogenicactivity. In particular, the invention relates to1,Z-methylene-B-nortestosterones optionally substituted at the6-position.

The compounds of the present invention are represented by the followingstructural formula:

The preferred reagent for this purpose is dimethyl sulfoxoniurnmethylide, which is formed in situ by reaction of dimethyl sulfoxidemethiodide with a strong base such as sodium hydride.

When W is 0H the starting materials of Formula II are obtained bydehydrogenating the corresponding B-norsteroid which is saturated at the1,2-position either microbiologically or chemically withdichlorodicyanobenzoquinone or selenium dioxide. A suitabledehydrogenating microorganism is Septomyxa afiim's NRRL 2746.

When W is C Cfiz CHz and R is methyl, the starting material is preparedas follows: 17a-methylandrost-S-ene-3[3,17fi-diol is acetylated to giveits 3,17-diacetate. Reaction with ozone, and then with zinc and aceticacid gives 3fl,17B-diacetoxy-17amethyl-5,6-secoandrostan-5-on-6-al.Oxidation with Jones reagent, followed by ring-closure with benzoylchloride in pyridine gives 3,8,17/3 diacetoxy 17amethyl-Sp-hydroxy-B-norandrostan-6-carboxylic acid 5,6-1actone.Reduction with lithium aluminum hydride gives a tetrol, and oxidationwith {N bromoacetamide gives 5,8,17B-dihydroxy-Gfl-hydroxymethyl 17ozmethyl-B-norandrostan-3- one. p-Toluenesulfonic acid dehydration of the6-hydroxymethyl group and -A-ring gives a 6-methylene-A -3-ketone andsubsequent reaction with dimethyl sulfoxonium methylide gives the6,6-ethylene compound. 1,2-dehydrogenation with a reagent such asdichlorodicyanobenzoquinone gives the required starting material. Use ofthe known 17a-ethyl starting material in the above reaction sequencegives the required 17u-ethyl compound. The 17DL'l1I1Sllbstitutedstarting material (R =H) is prepared as follows: S/E-acetoxy-SB-hydroxy17 oxo B norandrostan-G-carboxylic acid 5 ,6-lactone is reduced withlithium aluminum hydride to6,8-hydroxymethyl-B-norandrostane-318,56,17,8- triol. Oxidation withN-bromoacetamide gives the 3,17- dione and simultaneous dehydration ofthe fi-hydroxymethyl group and A-ring with p-toluenesulfonic acid gives6-methylene-B-norandrost-4-ene-3,17-dione. Reaction with dimethylsulfoxonium methylide, reduction of the 17- ketone, reoxidation of thesimultaneously reduced 3-ketone,, and dehydrogenation at the1,2-position gives the required starting material.

When W is the starting materials are prepared by dehydrogenating the6-methyl compounds saturated at the 1,2-position, which compounds areeither known or are themselves prepared by catalytic hydrogenation ofthe 6-methylene compounds.

The 1,2-rnethylene group of the product compounds of this invention isbelieved to possess the a-configuration. This configuration of the1,2-methylene group is accordingly designated by a dotted line. It is tobe understood, however, that the invention is intended to comprehend theactual compounds prepared by the processes herein disclosed, Whatevertheir configurations.

The compounds of the invention are antiandrogenic agents. They are to beused to alleviate conditions caused by hyperandrogenicity, such as acneand hirsutism. in standard antiandrogenic tests, they have been found topossess antiandrogenic activity when administered orally to rats atdoses of 50400 mg./kg. as sesame oil suspensions. They may be formulatedinto oral pharmaceutical compositions containing 5-1 00 mg. of activecompound or into a 1-5% topical cream by methods well known to skilledpharmaceutical chemists using standard pharmaceutical excipients.

The products of the invention may be converted into their obvious17-esters, including the acetate, propionate, butyrate,phenylpropionate, and cyclopentylpropionate by well-known acylationmethods. These esters, as obvious chemical variants of the 17-alcohols,are therefore considered equivalent thereto.

The following examples are intended to illustrate the preparation of thecompounds of the invention, but are not to be considered as limiting thescope thereof.

3 EXAMPLE l.-17B-hydroxy-17a-methyl-lu,2xmethylene-B-norandrost-4-en-3-one A solution of 1.0 g. of17fl-hydroxy-17a-methyl-B-norandrosta-1,4-dien-3-one in ml. of dimethylsulfoxide is treated in two portions with 25 ml. of a solution preparedfrom 4.2 g. of trimethyl sulfoxonium iodide, 50 m1. of dimethylsulfoxide, and 0.84 g. of 52% sodium hydride. The solution is stirred at25 for 3 hours, poured into water and the precipitate extracted intomethylene chloride. Drying and removal of solvent gives a crystallineresidue which is dissolved in 1:1 benzene-petroleum ether solution andfiltered through 30 g. of neutral alumina. The alumina is washed withmethylene chloride and the combined eluates are evaporated to acrystalline mass, then recrystallized from acetone-hexane solution togive the title product, M.P. 183. UV: e ,,=12,600. [a] =+17O.9 as 1.04%methanol solution.

When the corresponding 17ot-ethyl and 170L-11DS11bStituted compounds areused as starting materials in the above procedure,17(t-6thYl-17B-hydIOXy-10,20t-1'I16thYl6I18- B norandrost 4 en 3 one and17/3 hydroxy 10,20tmethylene B norandrost 4 en 3 one, respectively, areobtained.

EXAMPLE 2.6,6-ethylene- 17/3-hydroxy- 17 a-methyl-1a,2a-methylene-B-norandrost-4-en-3-one A solution of 0.5 g. of6,6-ethylene-17/3-hydroxy-17umethyl-B-norandrost-4-en-3-one in 25 ml. ofdioxane is heated with 0.5 g. of dichlorodicyanobenzoquinone and thesolution treated at reflux temperature for hours. The precipitatedhydroquinone is filtered, the solvent evaporated from the filtrate andthe residue then dissolved in benzene. The solution is filtered through15 g. of alumina, the solvent removed and the crystalline residuerecrystallized from acetone-hexane, then methanol-water solution to give6,6 ethylene 17B hydroxy 17oz methyl B- norandrosta 1,4 dien 3 one, M.P.182. UV: 624 62 4 9,4OO. [OC]D:69-6 as methanol solution.

A suspension of 11.7 g. of dimethyl sulfoxide methiodide in 100 ml. ofdimethyl sulfoxide is treated with 2.32 g. of 55% sodium hydride andvigorously stirred until the evolution of hydrogen has ceased. To 70 ml.of this solution 2.3 g. of6,6-ethylene-17B-hydroxy-17ot-methyl-B-norandrosta-l,4-dien-3-one isadded and the solution heated at 50 for 3.5 hours. The reaction solutionis cooled, poured into 800 ml. of water, filtered, washed with n-hexane,then water. The crude product is recrystallized from acetone-hexane andmethanol-water solutions to give6,6-ethylene-17fi-hydroxy-1aim-methylene- 17ot methyl B norandrost 4 en3 one, M.P. 203 UV: e ,,=12,600. [a] =+117.0 as 1.061% methanolsolution.

When 6,6 ethylene 175 hydroxy B norandrost- 4-en-3-one and 17a ethyl 6,6ethylene 17,6 hydroxy- B-norandrost-4-en-3-one are substituted for the17x-methyl starting material and the above dehydrogenation and methyleneaddition reactions are performed, 6,6-ethylene- 176 hydroxy 106,206methylene B norandrost 4 en- 3 one and 17cc ethyl 6,6 ethylene 17,8hydroxy- 1a,2a methylene B norandrost 4 en 3 one, respectively areobtained.

When 17p hydroxy 6 methyl B norandrost 4- en 3 -one, 17/8 hydroxy 6,17adimethyl B norandrost 4 en 3 one, and 1706 ethyl 17,8 hydroxy- 6 methylB norandrost 4 en 3 one are used as starting materials in the abovedehydrogenation and methylene addition reactions,l7fl-hydroxy-6-methyl-10:,2umethylene B norandrost 4 en 3 one, 1713hydroxy 6,17u dimethyl 10,2a methylene B norandrost 4 en 3 one, and 17aethyl 17B hydroxy- 6 methyl 1a,2u methylene 13 norandrost 4 en 3- cm ptive y, are obtained.

4 PREPARATION OF STARTING MATERIALS17fi-hydroxy-17u-methyl-B-norandrosta-1,4-dien-3-one A fermentationmedium composed of 10 liters of corn steep liquor adjusted to pH 6.3-6.5with sodium hydroxide solution is autoclaved for 2 hours at 15 p.s.i. at121. The medium is inoculated with a standard preparation of Septomyxaafiinis NRRL 2746. Fermentation growth is carried out for 48 hours withaeration at 3 liters of air per minute per 10 liters and impeller speedof 200 r.p.m.

Five grams of 17B-hydroxy-17a-methyl-B-norandrost4 en-3-one is dissolvedin 50 ml. of ethanol and added beneath the surface of the medium: 1 g.after 48 hours, 2 g. after 55 hours, 1 g. after 72 hours and 1 g. after78 hours.

After the transformation is complete, the mixture is centrifuged. Therecovered solids are extracted with ethanol and then ethanol-methylenechloride. The extracts are filtered and added to the clarified brothwhich is exhaustively extracted with methylene chloride. After drying,the extracts are evaporated in vacuo at 50.

The residue is taken into petroleum ether-benzene and chromatographedover alumina (Woelm, III). The fractions eluted with benzene petroleumether (1:1) through benzene-methylene chloride (1 :1) are combined andevaporated. The solid residue is recrystallized from acetonehexane togive 17fl-hydroxy-17ot-methyl-B-norandrosta- 1,4-en-3-one, M.P. 140-141after sublimation.

Use of 1749 hydroxy B norandrost 4 en 3 one and 17a ethyl 17B hydroxy Bnorandrost 4 en- 3-one as substrates in the above procedure givesl7fi-hydroxy B norandrosta 1,4 dien 3 one and17aethyl-17B-hydroxy-B-norandrosta-1,4-dien-3-one.

6,6-ethylene-17,8-hydroxy-17a-methyl-B- norandrost-4-en-3-0ne Reflux amixture of one part 17a-methylandrost-5-ene- 313,17fl-diol (60 grams)and 5 parts acetic anhydride (300 ml.) for one hour. Over the next hour,distill to remove 3 /2 parts (210 ml.) of distillateLCool somewhat andthen strip in vacuo to a crystalline slurry. Add water (ca. 2 parts, 120ml.) and stir for one hour to decompose anhydride. Dilute with water,stir for 15 minutes, filter and wash with water to obtain a slightlytacky crystalline solid. Transfer to a flask, add 2 parts (120 ml.)methanol and stir and reflux for 15-20 minutes. Allow to cool to roomtemperature, filter, wash with 1-1.5 parts (60-90 ml.) methanol and dry.Weight of the 3,17-diacetate is 64 grams, M.P. 137-139, which is ofsufiicient purity for the next reaction.

Dissolve 24 grams of l7a-methylandrost-5-ene-3fl,175- diol,3,17-diacetate in 30 parts (720 ml.) methylene chloride containing 1%methanol (7.2 ml.). Cool in acetone-Dry Ice bath and ozonize withoxygen-ozone mixture until solution takes on a permanent blue colorindicating presence of excess ozone. Transfer to an ice-salt bath andadd 3 parts (72 g.) of zinc dust. To the stirred mixture slowly add 4parts (96 ml.) of acetic acid. Stir in ice bath for one hour, thenremove bath and stir for one hour at room temperature. Filter off zincand wash well with methylene chloride. Wash solution with water,bicarbonate, and water. Dry solution over Na SO then remove solvent invacuo and crystallize the residue from ether-petroleum ether to obtain17 grams of 3p, 17,8-diacetoxy 17cc methyl-5,6-secondrostan-5-on-6-al,M. P. 118-120". Recrystallized product melts at 122-124. The motherliquor contains additional aldehyde which can be processed further .tothe lactone. Unless crystalline aldehyde is desired the total residuefrom the ozonization is oxidized to the carboxylic acid and convertedinto the lactone without isolation. For this the CH Cl solution isconcentrated .to 4 parts by volume (ca. ml.).

Dilute solution of 6-aldehyde in 4 parts of CH CI with 8 parts (200 ml.)of acetone. Stir and cool in ice ba h o 35. Over a 3 to 5 minute periodadd 0.8 part (19.2

ml.) of Jones oxidizing reagent whereby temperature may rise to 1012.After the addition, stir for 10-15 minutes more, then add 800 ml. water.Extract with CH Cl and wash with water to remove Cr salts. Concentratein partial vacuum to remove solvent and obtain an oily residue of the6-oic acid. Dissolve residue in parts (120 ml.) pyridine and stir andcool in an ice bath. Add dropwise 1.1 parts (26.4 ml.) of benzoylchloride, then stir in bath for a half hour. Remove from bath and stirat room temperature for 3 hours (or longer). Add 1 part Water (24 ml.)and stir for one hour at room temperature. Dilute with water and extractwith benzeneether. Wash solution successively with water, dilute HCl,water, dilute NaOH, and water. Concentration in vaco to a crystallineresidue, add methanol, stir and concenrate to remove benzene, ending upin methanol at a volume of 2 parts (ca. 50 ml.). Cool, filter, and washwith MeOH. Yield of 36,176 diacetoxy-17u-methyl-5fi hydroxy-B-norandrostan-6-carboxylic acid 5,6-lactone is 18.5-19.2 g., M.P.167-169.

Under nitrogen add portionwise 6.5 grams of LiAlH to a stirred mixtureof 13 grams of the lactone in 500 ml. of tetrahydrofuran. After theaddition, reflux for 2 hours, then cautiously add mixture of 25 ml.water and 50 ml. tetrahydrofuran to decompose excess reagent. Refluxgently for a half hour, filter, and wash with tetrahydrofuran. Removesolvent in vacuo to a mostly crystalline residue which is taken up inmethanol and benzene. Concentrate to remove methanol and to crystallizein benzene. Allow to cool to room temperature, filter, wash with benzeneto obtain6/3-hydroxymethyl-17a-methyl-B-norandrostane-3B,5fi,17fi-triol, M. P.200-210, which is of sufficient purity for the next oxidation step. Thetetrol can be recrystallized from MeOH, M.P. 217-219.

Stir a mixture of grams of the tetrol in 200 ml. t-butyl alcohol and 200ml. methanol. Add 8 ml. pyridine, followed by 6.6 gramsN-bromoacetamide. Stir in dark overnight. Add 60 ml. water and 3.3 gramsNaHSO and stir to decolorize. Concentrate in vacuo to remove solventsand obtain a thick slurry which is then diluted with waer to 200-250 ml.volume. Stir for one hour and filter and wash with water. Weight of55,17,8-dihydroxy-6B- hydroxymethyl 17a-methyl-Bmorandrostan-3-one afterdrying is 8.0-8.3 grams, M.P. 170-175.

To a stirred mixture of 10 grams of the triol ketone in 900 ml. benzeneand 100 ml. dioxane maintained at 40 by means of a bath, add 1.25 g. ofp-toluenesulfonic acid. Stir for several hours at 40, then slowly raisetemperature to 5055. Continue reaction to completion as determined byTLC. so as to dehydrate tto the 3-keto-A and to the 6-methylene withminimum loss of the 17- hydroxy group. This will require 6-8 hours time,during which the solution gradually develop color. At end of reaction,wash benzene solution with water, Na CO and water. Dry over Na SO andremove solvent in vacuo to a mostly crystalline residue which isdigested in ether, filtered, and washed with ether to give the6-methylene- 17 3 hydroxy-17a-methyl-B-norandrost-4-en-3-one, M.P.162-165 Under nitrogen, add in portions 0.65 gram of 55% NaH to 3.3grams of dimethyl sulfoxide methiodide in 50 ml. dimethyl sulfoxide.Stir for 20 minutes to obtain complete solution, then add 3.0 g. of the6-methylene compound. Stir at room temperature for one hour, then anadditonal hour at 45-50". Pour reaction mixture into ice and water anddilute to 400 ml. Filter and wash with water. Dissolve wet cake inbenzene with aid of MeOH) and dilute with ether. Wash solutionsuccessively with water, dilute NaOH, and water, discarding small cuffsobtained. Dry over Na SO remove solvent in vacuo to a crystallineresidue which is then crystallized from acetone-hexane or from ethylacetate. The product, 6,6- ethylene 17,3hydroxy-17a-methyl-B-norandrost-4-en-3- one, melts at 187-189.

The 170: ethyl-6,6-ethylene-17fl-hydroxy-B-norandrost- 4-en-3-one isprepared in the same manner from the known17/8-ethylandrost-5-ene-3p,17,8-diol.

6,6-ethylene-17B-hydroxy-B-norandrost-4-en-3 -one To a stirredsuspension of 20 g. of lithium aluminum hydride in 700 ml. oftetrahydrofuran under a nitrogen atmosphere is slowly added 30 g. of3fl-acetoxy-5fl-hydroxy-17-oxo-B-norandrostan-6-carboxylic acid5,6-lactone [Tanabe et al., Chem. Pharm. Bull., 9, 12 (1961)] in 250 ml.of tetrahydrofuran. After addition is complete, the reaction mixture isstirred at reflux for 2 hours. After cooling with an ice bath, thereaction mixture i slowly treated with m1. of water in ml. oftetrahydrofuran. The resulting white precipitate is removed byfiltration and washed with fresh tetrahydrofuran. Evaporation of thefiltrate gives a residue of Gfl-hydroxymethyl-B-norandrostane-3B,5,8,17fl-triol, M.P. 166170 after crystallization fromethyl acetate.

To a stirred solution of 60 g. of N-bromoacetamide in 1 l. of methanol,protected from direct light, is added 18 ml. of pyridine, 60 ml. ofwater and 18 g. of the above tetrol. After stirring at room temperaturefor 16 hours, 40 g. of solid Na SO followed by 400 ml. of water isadded, stirring is continued for a few minutes, and finally the reactionmixture is concentrated to 500 ml. at reduced pressure. The warm mixtureis diluted with 1.5 l. of water, cooled, and filtered to givecrystalline 6p hydroxymethyl-5/3-hydroxy-B-norandrostane 3,17- dione,M.P. 178-180 after recrystallization from ethyl acetate.

A solution of 5 g. of the above diketone in 500 ml. of benzenecontaining 0.5 g. of p-toluenesulfonic acid is stirred at roomtemperature under nitrogen for 10 hours. The reaction mixture is washedwith dilute aqueous sodium carbonate solution, dried, and evaporated toa residue of 6-methylene-B-norandrost-4-ene-3,l7 dione, which afterrecrystallization from methanol has M.P. 143-148, UV: max. 283 my. (615,500).

To a stirred solution of 9.3 g. of dimethyl sulfoxide methiodide in 90ml. of dimethyl sulfoxide under a nitrogen atmosphere is slowly added1.6 g. of NaH as a 55% mineral oil suspension. After stirring for 10minutes, 10 g. of the 6-methylene compound in ml. of dimethyl sulfoxideis added. The reaction mixture is stirred for 30 minutes at roomtemperature and then at 60 for 45 minutes. It is then poured into Waterand extracted with benzene. Evaporation of the dried benzene extractsgives a residue of 6,6-ethylene-B-norandrost-4-ene-3,17-dione, M.P.205-209, UV: A max. 262 m (6 15,900) after recrystallization fromacetone-hexane.

To a stirred suspension of 1.7 g. of LiAlH in 85 ml. of ether under anitrogen atmosphere is slowly added 6.8 g. of6,6-ethylene-B-norandrost-4-ene-3,17-dione in 110 ml. oftetrahydrofuran. After addition is complete, the reaction mixture isrefluxed for 2 hours, cooled, treated with 7 ml. of Water in 20 ml. oftetrahydrofuran, filtered and the filtrate evaporated to a crude residueof 6,6- ethylene-B-norandrost-4-ene-3[3,17,8-diol. Without furtherpurification the crude diol is dissolved in 100 ml. of dioxane andtreated with 5.7 g. of 2,3-dichloro-5,6-dicyanobenzoquinone in 70 ml. ofdioxane. After 5 hours at room temperature, the reaction mixture isfiltered and the filtrate is evaporated to a residue which is dissolvedin methylene chloride-benzene (1:1) and chromatographed on 200 g. ofalumina (activity III, Woelm). Elution with methylene chloride-benzene(1:1) and methylene chloride gives6,6-ethylene-17/3-hydroxy-B-norandrost-4-en-3-one, M.P. -177" afterrecrystallization from ethyl acetate.

17 fl-hydroxy-6, 17a-dimethyl-B-norandrost-4-en-3 -one To a Parrhydrogenation bottle is added 300 mg. of 5% Pd/CaCO 75 ml. of methanol,4.5 ml. of pyridine, and a solution of 3.0 g. of17,8-hydroxy-17a-methyl-6- methylene-B- norandrost-4-en-3-one in 75 ml.of methylene chloride. The hydrogenation is conducted at 20 p.s.i.

7 for /2 hour by which time one mole of hydrogen has been absorbed. Thecatalyst is filtered off and the filtrate evaporated in vacuo to an oilwhich is a mixture of the 604 and 6,8-methyl isomers. The isomers can beseparated by crystallization or by neutral alumina chromatography.Crystallization of the oil from ether gives the less soluble 60t-iSOm6I,M.P. 178180, and crystallization of the remaining oil fromether-petroleum ether gives the more soluble 6fi-isomer, M.P. 137140;recrystallized 140- 141. By chromatography, the 6fi-isomer is elutedfrom the alumina column with 1:1 benzene-petroleum ether and the60t-iSOm6r with benzene. Mixtures of the isomers are isomerized to the6,8-isomer by treatment with alcoholic alkali.

The 17a-ethyl 175-hydroxy-6-methyl-B-norandrost-4- en-3-one startingmaterial is prepared in the same manner from the corresponding17tx-ethyl-6-methylene compound.

The 175 hydroxy-6-methyl-B-norandrost 4-en-3-one starting material is aknown compound.

I claim:

1. A compound of the formula where W is 1r 0 r1 0113MB orn and R ishydrogen, methyl, or ethyl.

2. A compound as claimed in claim 1. Where W is CHzCH2 3. A compound asclaimed in claim 2, where R is methyl, being the compound6,6-ethylene-17B-hydroxy-17a-methyl-1(1,20:-methylene-B-norandrost-4-en-3-one.

4. A compound as claimed in claim 1, being the compound 17/3-hydroxy6,17a-dimethyl-1a,2ot-methylene-B- norandrost-4-en-3-one.

References Cited UNITED STATES PATENTS 3,356,716 12/1967 Holden 260488FOREIGN PATENTS 9/1965 Netherlands.

OTHER REFERENCES Saunders et al.: Chem. Abst., vol. 61, col. 8594B,(1964).

Fieser et al.: Steroids, p. 519 (1959).

LEON ZITVER, Primary Examiner.

M. M. JACOB, Assistant Examiner.

Us 01. X.R.

